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Blood, 1 May 2001, Vol. 97, No. 9, pp. 2611-2617
HEMATOPOIESIS
A novel myeloid-restricted zebrafish CCAAT/enhancer-binding
protein with a potent transcriptional activation domain
Susan E. Lyons,
Bixiong C. Shue,
Andrew C. Oates,
Leonard I. Zon, and
P.
Paul Liu
From the National Human Genome Research Institute,
National Institutes of Health, Bethesda, MD, and the Howard Hughes
Medical Institute, Children's Hospital, Harvard Medical School,
Boston, MA.
The CCAAT/enhancer-binding protein (C/EBP) family consists of
transcription factors essential for hematopoiesis. The defining feature
of the C/EBPs is a highly conserved carboxy-terminal bZIP domain that
is necessary and sufficient for dimerization and DNA binding, whereas
their amino-terminal domains are unique. This study reports a novel
c/ebp gene (c/ebp1) from zebrafish that encodes
a protein homologous to mammalian C/EBPs within the bZIP domain, but
with an amino terminus lacking homology to any C/EBP or to any known
sequence. In zebrafish embryos, c/ebp1 expression was
initially observed in cells within the yolk sac circulation valley at
approximately the 16-to 18-somite stage, and at 24 hours postfertilization (hpf), also in circulating cells. Most
c/ebp1+ cells also expressed a known early
macrophage marker, leukocyte-specific plastin (l-plastin).
Expression of both markers was lost in cloche, a mutant
affecting hematopoiesis at the level of the hemangioblast. Expression
of both markers was retained in m683 and
spadetail, mutants affecting erythropoiesis, but not
myelopoiesis. Further, c/ebp1 expression was lost in a
mutant with defective myelopoiesis, but intact erythropoiesis. These
data suggest that c/ebp1 is expressed exclusively in
myeloid cells. In electrophoretic mobility shift assays, c/ebp1 was
able to bind a C/EBP consensus DNA site. Further, a chimeric protein
containing the amino-terminal domain of c/ebp1 fused to the DNA-binding
domain of GAL4 induced a GAL4 reporter 4000-fold in NIH3T3 cells. These
results suggest that c/ebp1 is a novel member of the C/EBP family that
may function as a potent transcriptional activator in myeloid cells.
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